Methods for amplifying and detecting a predetermined target nucleic acid in a biological specimen are known, and can be accomplished even when there is a mismatch in a single position between a primer and the target nucleic acid. The mismatch is located at or near the 3 min end of the primer. Disclosed herein is a method for overcoming such a mismatch, and is particularly useful for detection of a nucleic acid sequence which is not fully known, or where there is considerable heterogeneity in DNA target from patient samples. The method is characterized by using a primer having a nucleotide with a thymine base at the position of the mismatch, the primer being used to prime the target and to form primer extension products.
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